Method of modulating neovascularization

Abstract

The invention provides a method of inhibiting neovascularization in a subject. The method comprises administering to the subject an agent that interferes with fibronectin (Fn) matrix assembly in an amount effective to inhibit neovascularization. The invention also provides a method of identifying an agent that inhibits neovascularization. The method comprises detecting fibronectin (Fn) matrix assembly by stimulated endothelial cells cultured in three-dimensional culture gel in the presence and absence of an agent. A decrease in Fn matrix assembly in the presence of the agent compared to Fn matrix assembly in the absence of the agent is indicative of an agent that inhibits neovascularization. Alternatively, the method of identifying an agent that inhibits neovascularization comprises detecting changes in nuclear architecture in stimulated endothelial cells cultured in three-dimensional culture gel in the presence and absence of an agent. A reduction in nuclear architecture organization identifies an agent that inhibits neovascularization.

Description

CROSS REFERENCE TO RELATED APPLICATION[0001]This application claims priority to U.S. Provisional Patent Application No. 61 / 043,610, filed Apr. 9, 2008.GRANT FUNDING DISCLOSURE[0002]This invention was made with government support under grant number R01 CA88308, awarded by the National Institutes of Health. The government has certain rights in the invention.FIELD OF THE INVENTION[0003]The invention relates to materials and methods of modulating neovascularization.BACKGROUND OF THE INVENTION[0004]Neovascularization, or the formation of new blood vessels, is a highly complex and tightly regulated biological process. Neovascularization begins with the enzymatic breakdown of the basement membrane of a blood vessel. Endothelial cells migrate through the area of degradation, invade the surrounding extracellular matrix, and proliferate to form an elongated column of cells. A lumen forms within the solid cell column upon differentiation of endothelial cells and the basement membrane is subseq...

AI Technical Summary

Benefits of technology

[0029]“Derivatives” include agents that have been chemically modified in some manner distinct from insertion, deletion, or substitution variants. In this regard, the agent is chemically bonded with polymers, lipids, other organic moieties, and / or inorganic moieties. Derivatives are prepared in some embodiments to increase solubility, absorption, circulating half-life, or targeting to particular cells, tissues, or organs. Chemical modification also may eliminate or attenuate any undesirable side effect of the agent, such as immunogenicity. In this regard, agents covalently modified to include one or more water soluble polymer attachments, such as polyethylene glycol, polyoxyethylene glycol, or polypropylene glycol are contemplated herein (U.S. Pat. Nos. 4,640,835; 4,496,689; 4,301,144; 4,670,417; 4,791,192; and 4,179,337). Still other useful polymers known in the art include monomethoxy-polyethylene glycol, dextran, cellulose, or other carbohydrate based polymers, poly-(N-vinyl pyrrolidone)-polyethylene glycol, propylene glycol homopolymers, a polypropylene oxide / ethylene oxide co-polymer, polyoxyethylated polyols (e.g., glycerol) and polyvinyl alcohol, as well as mixtures of any of the foregoing.

[0068]Prior to embedding in the 3-D fibrin matrix, endothelial cells were ballistically microinjected with 100 nm polystyrene beads to circumvent the endocytic pathway and subsequent directed motion of the beads. After 3 days of incubation, the beads dispersed in the cytoplasm and their centroids were tracked with high spatial and temporal resolution using fluorescence microscopy. Relative to control endothelial cells, the mean square displacement (MSD) of the beads was significantly increased in cells treated with the FUD peptide, indicating a significant relative cytoplasmic softening compared to that of cells where Fn matrix assembly is intact. Elastic moduli, which quantify the local resistance of the cytoplasm against small random forces acting on the surface of the beads, were derived from MSD curves to quantify cellular mechanical properties. The elastic modulus of the cytoplasm of FUD-treated cells is significantly lower than that of control cells (P<0.001), indicating a pronounced defect in internal stiffness and the cell's ability to sense a sufficiently rigid substratum.

Problems solved by technology

When vascularization is not stringently controlled, serious pathologies can result.

However, a major hurdle in treating or preventing angiogenesis is targeting processes uniquely associated with unwanted neovascularization to avoid side effects.